The G-switch™ Direct line of small G-protein modulators have been developed with an emphasis on creating highly potent cell permeable reagents that directly target the Rho family of small G-proteins. Our Direct Activator reagents are based on the catalytic domain of the bacterial CNF toxins, which are covalently attached to a proprietary cell penetrating moiety. The Rho/Rac/Cdc42 Activator I (Cat# CN04) enters the cell and activates Rho GTPase isoforms by deamidating glutamine-63 of Rho and glutamine-61 of Rac and Cdc42 in their respective Switch II regions (1,2). This modification converts glutamine-63 to glutamate, which blocks intrinsic and GAP stimulated GTPase activity resulting in constitutively active Rho (3). The Rho/Rac/Cdc42 Direct Activator robustly increases the level of GTP bound RhoA, Rac1 and Cdc42 within 2-4 h after addition to the culture medium and thus provides a more rapid alternative to transfection based methods for introducing activators like GEFs into cells. Moreover, the targeted action of the Rho Direct Activator makes it far more attractive tool for the study of Rho GTPase signaling than classic indirect activators (e.g. LPA, EGF, Bradykinin and Sphingosine-1-phosphate) that concomitantly activate other signaling pathways (e.g. Ras, PI3K and PLC).
